Departamento de Química Analítica, Química Física e Ingeniería Química, Universidad de Alcalá, Ctra. Madrid-Barcelona Km. 33.600, 28871, Alcalá de Henares (Madrid), Spain.
Departamento de Química Analítica, Química Física e Ingeniería Química, Universidad de Alcalá, Ctra. Madrid-Barcelona Km. 33.600, 28871, Alcalá de Henares (Madrid), Spain; Madrid Institute for Advanced Studies of Water (IMDEA Agua), Parque Científico Tecnológico, E-28805, Alcalá de Henares (Madrid), Spain.
J Chromatogr A. 2021 Feb 22;1639:461919. doi: 10.1016/j.chroma.2021.461919. Epub 2021 Jan 20.
The first CE methodology enabling the enantiomeric separation of panthenol was developed in this work. Electrokinetic chromatography with cyclodextrins (CD-EKC) was the CE mode employed for this purpose. The effect of different experimental variables such as the nature and concentration of the cyclodextrin, the temperature and the separation voltage was investigated. The best enantiomeric separation was obtained with 25 mM (2-carboxyethyl)-β-CD (CE-β-CD) in 100 mM borate buffer (pH 9.0), with a separation voltage of 30 kV and a temperature of 30 °C. Under these conditions, an enantiomeric resolution of 2.0 in an analysis time of 4.2 min was obtained, being the biologically active enantiomer d-panthenol (dexpanthenol) the second-migrating enantiomer. The analytical characteristics of the method were evaluated in terms of precision, accuracy, selectivity, linearity, LOD, and LOQ, showing a good performance for the quantitation of dexpanthenol in cosmetic and pharmaceutical formulations. The enantiomeric impurity (L-panthenol) could be detected at a 0.1% level with respect to the majority enantiomer, allowing to accomplish the requirements of the ICH guidelines. The method was also successfully applied to study the stability of panthenol under abiotic and biotic conditions and its toxicity on non-target organisms (the aquatic plant Spirodela polyrhiza).
本文开发了第一种能够对泛醇对映体进行拆分的 CE 方法。为此目的,采用了环糊精电动色谱(CD-EKC)作为 CE 模式。研究了不同实验变量的影响,例如环糊精的性质和浓度、温度和分离电压。在 25 mM(2-羧乙基)-β-CD(CE-β-CD)和 100 mM 硼酸盐缓冲液(pH 9.0)中,在 30 kV 的分离电压和 30°C 的温度下,获得了最佳的对映体分离。在此条件下,分析时间为 4.2 分钟时,对映体分辨率为 2.0,具有生物活性的对映体 d-泛醇(dexpanthenol)为第二迁移对映体。该方法的分析特性通过精密度、准确度、选择性、线性、LOD 和 LOQ 进行了评估,在化妆品和药物制剂中定量测定 dexpanthenol 时表现出良好的性能。可以检测到相对于主要对映体的 0.1%水平的对映体杂质(L-泛醇),从而满足 ICH 指南的要求。该方法还成功地应用于研究泛醇在非生物和生物条件下的稳定性及其对非目标生物(水生植物水蕹)的毒性。